CONTAINER FOR DETERMINING OF ANALYTES AND/OR CHEMICO-PHYSICAL PARAMETERS, AS WELL AS DETERMINING OF URINARY SEDIMENT, IN URINE; AND METHOD OF FULL URINE ANALYSIS USING THIS CONTAINER

Abstract

The present invention relates to a container (1) apt to contain urine, for the determination of analytes and/or chemico-physical parameters, and of urinary sediment, via an automated analyzer, the container comprising a) a containment region (9) having an opening (3) with a tightenable and pierceable closure element (4); b) a reading region (5) with one or more colorimetric reaction pads (6) made integral with an inner wall of the container; c) an impermeable and pierceable separation septum (10) positioned between the containment region (9) and the reading region (5). The present invention also relates to a method of running a full test of urine on an automated analyzer, including physical-chemical examination and urine sediment, which comprises the steps of a) filling the above-mentioned container (1) from a primary collection container by suction regulated from pre-calibrated vacuum; b) loading the container (1) on the automatic analyzer; c) piercing the closure element (4) with a sampler and mixer needle (20); d) mixing the sample in the containment region (9) and withdrawing some urine to be transferred to the analyzer for the automatic reading of the urinary sediment e) piercing the septum (10) and retracting the sampler needle (20) such that urine flows into the reading region (5) f) waiting for a specified time for the generation of color reactions; g) performing an automatic reading of the colorimetric reactions using a sensor integrated in the analyzer.

Claims

1. Container adapted to running a standard full urine test comprising determination of analytes and/or chemico-physical parameters and of urinary sediment on native urine via an automated analyzer in a closed mode, comprising: a) a containment region having an opening with a tighten and pierceable and recloseable closure element, suitable for sampling by a needle in automated and closed mode; b) a reading region bringing one or more reaction pads made integral to an inner wall of the container, each reaction pad being adapted to absorb a pre-established amount of urine and to give rise to a colorimetric reaction specific for a respective analyte, parameter and/or reference; and c) a separation septum, impermeable and pierceable, positioned between the containment region and the reading region.

2. Container according to claim 1, wherein the containment region has a precalibrated vacuum suitable for the recall of a quantity of urine from a primary container of collected sample.

3. Container according to claim 1, wherein said separation septum is made, at least in part, of aluminum.

4. Container according to claim 3, wherein said separation septum comprises a peripheral region (11) in plastic material and a pierceable central region in aluminum.

5. Container according to claim 1, wherein said inner wall of the main body has at least a recess zone adapted to contain a respective pad for its whole thickness.

6. Container according to claim 1, wherein one or more of said pads are made on a strip to be fixed at an inner wall of said reading region.

7. Container according to claim 6, wherein said inner wall of the reading region has a recess zone adapted to contain said strip for its whole thickness.

8. Container according to claim 1, wherein said containment region and said reading region have at least a portion of plane wall, said at least a portion of plane wall, being destined to house said reaction pads.

9. Container according to claim 1, wherein each reaction pad is associated with a code identifying a correspondent analyte and/or parameter, adapted to be recognized by an optical reader integrated in the automated analyzer.

10. Container according to claim 1, wherein the zones between different reaction pads present in the reading region are made opaque.

11. Container according to claim 1, comprising a protective non-permeable film, made adherent at its back to said reaction pads on their side faced to the inner wall of the container in the reading region, and having a calibrated hole at each of said pads allowing contact with a correct amount of urine.

12. Method to run a standard full test of urine on an automated analyzer, including physical-chemical examination and urine sediment on original (native) urine via an automated analyzer in closed mode, which comprises the following: a) filling a container according to claim 1 with a urine sample, from a primary collection container by suction regulated from pre-calibrated vacuum, in closed mode from the outside; b) loading the container on the automatic analyzer; c) piercing the pierceable and reclosable closure element with a sampler and mixer needle so as to inset the needle in the containment region according to a closed mode; d) mixing the sample present in the containment region and withdraw in a closed mode a predetermined amount of original (native) urine; e) transferring such predetermined amount to the analyzer for the automatic reading of urinary sediment; f) piercing the pireceable and recloseable closure element and the septum, retracting the sampler needle, in such a way that the urine flows in the reading region coming into contact with the reaction pads; g) waiting for a specified time for generation of color reactions; and h) performing an automatic reading of the colorimetric reaction using a sensor integrated into the analyzer.

13. Method according to claim 12, wherein said reading step provides for a rotation of the container along its longitudinal axis.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0056] The advantages arising from the present invention, together with the characteristics and conditions of employment of the given device, will become clear from the following detailed description, relating to its embodiments, suitable to the purposes above mentioned, and presented for illustrative but not limitative purposes.

[0057] We will refer to the figures of the attached drawings, where:

[0058] FIG. 1 is a perspective view of a container according to an embodiment of the present invention;

[0059] FIGS. 2A, 2B and 2C show, as example, possible variations of embodiments of the container according to the present invention;

[0060] FIGS. 3A to 3D illustrate how to use a container according to further embodiments of the present invention;

[0061] FIGS. 3E and 3F show a possible embodiment of a separation septum for a container according to the present invention;

[0062] FIGS. 4A and 4B show a container according to a further embodiment of the present invention;

[0063] FIGS. 5A and 5B show a container according to a further embodiment of the present invention;

[0064] FIGS. 6A and 6B show, as example, possible forms of the reaction pad according to the present invention.

[0065] The present invention will be described below with reference to Figures above mentioned.

[0066] With reference to FIG. 1, this shows a container for urine samples according to one embodiment of the present invention.

[0067] For the purposes of this description, the container 1 will always be described as a test tube, thus having a body 2 with a tubular shape, elongated and with a main body including a containment region 9 of the urinary material, having an opening 3 at its upper end and an underlying reading region 5 in which are allocated one or more reaction pads.

[0068] The container includes a pre-calibrated vacuum in its interior, in correspondence of the containment region, so as to effectively respond to the usual safety requirements.

[0069] However, it has to be clarified that such a tubular shape of the test tube is not a limiting feature of the present invention. There may be in fact other forms responding to the same inventive concept.

[0070] The container includes a closure element 4 to seal in a removable manner the opening 3. The closure element 4 may be applied to the container through a screw system, pressure, or other equivalent system. The closure element 4 will have to be made by material pierceable by a sampler/mixer needle 20 and will have to ensure the tightness with the external environment even after the said needle 20 will be spilled from the element 4.

[0071] As already indicated, the container 1 according to the present invention provides a reading region 5. Such reading region 5 holds one or more reaction pads 6, made integral to an inner wall of the main body 2.

[0072] Each reaction pad is able to absorb the fluid and to result in a colorimetric reaction specifically for a respective analyte and/or measurement of interest.

[0073] One of the advantages of such a realization is that it is not longer necessary to pick up and carry various amounts of urine outside the device to perform the biochemical reaction on a specific automated analyzer.

[0074] For the realization of the reaction pad, and of the reactive components specific for the different analytes and/or parameters to be determined, see the following description.

[0075] The following FIGS. 2A to 2C illustrate several variants relating to the allocation mode of the reaction pads 6 on the inner wall of the container, in correspondence with the reading region.

[0076] In particular, the first possibility is illustrated in FIG. 2A, in which the reaction pads (one or more of them) are based on a strip 7 which then is fixed to an inner wall of the main body, in correspondence with the reading region 5.

[0077] Advantageously, this strip 7 is made of non-absorbent material, so as not to interfere with the imbibition of the reaction pad. Moreover, it can advantageously be done in order to be opaque in the space between a pad and the other, so as to reduce or eliminate possible interferences during the colorimetric reading of the results.

[0078] In FIG. 2B a further variant is shown, with the inner wall of the main body having a recess adapted to contain the area of the strip 7 along its entire thickness.

[0079] FIG. 2C illustrates a further variant, with the inner wall of the main body presenting, for each reaction pad, a corresponding area of recess adapted to contain the pad for its entire thickness.

[0080] The advantage of the embodiments illustrated in FIGS. 2B and 2C (with particular reference to the presence of a recess) is constituted by a greater stability of placement of the reaction pad, imprisoned within a specific housing, in order to avoid any defects in adhesion and/or welding of the same in the test tube itself.

[0081] In all cases above described, it is advantageous that the areas of the main body between the distinct reaction pads are opaque, in order to reduce or eliminate possible interference during the colorimetric reading of the results.

[0082] According to a possible embodiment of the invention, to the rear of each individual pad of reaction, on the side facing the inside of the container, an insulating protection film 30 can advantageously be provided, having a calibrated hole 31 to allow contact with a correct urine amount, thus avoiding an excessive imbibition of the pad with the same urine.

[0083] One of the critical aspects in this type of analysis is definitely the reaction time, which, as already specified, is particularly short (60-120).

[0084] It follows that, in the known procedure, the urine collection should take place in suitable containers, and that the specific colorimetric reactions occur subsequently, or following manual insertion of the strips in the urine sample or following automatic withdrawal of small amounts of urine liquid that are distributed on other reaction elements, external to the device containing the sample.

[0085] According to an embodiment, the present invention addresses and solves this problem too.

[0086] In particular, the figures from 3A to 3D relate to this embodiment.

[0087] With reference to FIG. 3A, the container 1 may include a separation septum 10, impermeable and pierceable, positioned between the containment region and the reading one.

[0088] The septum 10 is a separation surface able to divide into two separate compartments the inside part of the test tube itself: the collection or containment compartment 9 and the reading region 5.

[0089] The compartment 9 is designed for the initial collection of the urine to be analyzed, without this being in contact with the reaction pads which instead are located in the reading region 5.

[0090] Of course it is to be considered that the septum 10 will be made in suitable materials, as not to contaminate the possible biological matrix.

[0091] For example, the separation septum can be made entirely in the form of an aluminum sheet, easily drillable.

[0092] Alternatively, as visible in FIGS. 3E and 3F, the separation septum may comprise a peripheral region 11 of plastic material and a central region (12), made of aluminum and pierceable.

[0093] As illustrated in FIG. 3B, in the use of the container 1 on an automatic analyzer, a sampler and mixer needle 20 can be used to perforate the closure element 4 and mix the urine present in the containment region 9.

[0094] Then, withdrawing via the sampler needle 20 a predetermined quantity of urine, this can be dedicated to reading of the urinary sediment.

[0095] Subsequently, as shown in FIG. 3C, always through the sampler needle 20, the perforation of the septum 10 it can be carried out.

[0096] As illustrated in FIG. 3B, the drilling of such a surface, within an analyzer, can take place e.g. by a needle sampler 20, or with any other equivalent device.

[0097] The drilling of the septum 10 then allows the urinary liquid to flow in the reading region of the test tube, to soak the reaction pad and therefore to result in the colorimetric reactions within the correct timing (FIG. 3D).

[0098] As already indicated, it is not strictly necessary that the container according to the present invention presents in the form of a classic test tube.

[0099] In particular, the container may not have an exactly cylindrical shape for the whole or part of its height. For example, some portions of the wall may be flat, so as to enable the sensor of the analyzer for a better reading of the colorimetric reactions from the reaction pad.

[0100] Therefore, there may be provided, for example, shapes with different section for the test tube or for parts of it.

[0101] Based on the location of the reaction pad and the conformation of the tube, it may be provided for an automatic rotation of the tube itself during the reading of the analyzer.

[0102] FIGS. 4A and 4B show, as example, a possible shape for the container according to the present invention.

[0103] In this case, the container has a tubular shape, in cross section, has two opposite flat faces 15, 16.

[0104] On each of these faces, the reaction pads 6 are placed, as so far described.

[0105] FIGS. 5A and 5B, however, relate to an embodiment according to the test tube with a single flat surface 17, inside which the reaction pads are placed, for example on two columns.

[0106] Advantageously, and for each of the embodiments described so far, each reaction pad is associated to an identification barcode not alphanumeric, related to the corresponding analyte determined by it.

[0107] FIGS. 6A and 6B relate to the possible implementations of identification codes associated to the reaction pads.

[0108] In particular, each reagent pad may be marked, on the top or bottom side, with an identification code (e.g. barcode or QRcode) of the pad itself, and/or of the single reaction, that can be detected by automatic sensors.

[0109] The codes may be prepared in common assembly with each reaction pad or on adherent strip inside the test tube, so that each code matches a single reference pad.

[0110] It is to be considered that the shape of the reaction pad can be different (square, round, rectangular, etc.) depending on the specific embodiment of the container, so as to allow a better stability in the housing, and an optimal reading surface of the sensor.

[0111] As example, FIG. 6B shows a square reaction pad, with an associated barcode right adjacent to one of the four sides.

[0112] Moreover, according to possible alternative embodiments, it can be envisaged that one or more of the reaction pads have the function of reactive control pads, to be used as reference measurement during the automatic reading.

[0113] This spot has the task of indicating, whereas the color change of the colorimetric range exceeds the maximum tolerated, the excessive prolongation of the contact time between the reaction pad and the urine, and therefore the automatic reading of the test tube (or the possible re-reading by subsequent passage on the analyzer) would be wrong, thus not being possible its validations.

[0114] Advantageously it can be inserted, on the inside part of the tube, even a spot capable of detecting the indices of interference with the optical reading.

[0115] Moreover, the codes 21, 22, indicating the start and end of the automatic reading, can be inserted.

[0116] For completeness, just as example, following there is a brief description of general chemical principles that determine the reactions between the used substrates and the main parameters tested in the urine:

[0117] Bilirubin

[0118] The coupling of bilirubin with a diazonium salt in an acidic environment results in a red azo coloring agent.

[0119] In the range of colors the following values are reported: 0 (negative), 1 (+), 2 (++), 4 (+++) mg/dl or resp. 0 (negative), 17 (+), 35 (++), 70 (+++) mol/l.

[0120] Bilirubin concentrations are reported starting from 0.5-1 mg/dl.

[0121] Proteins

[0122] The test is based on the principle of the protein error of a pH indicator. The test is primarily reactive with albumin.

[0123] In the range of colors the following concentrations of albumin are reported: negative, 30, 100, and 500 mg/dl and resp. negative, 0.3, 1.0 and 5.0 g/I. The concentrations of albumin are reported starting from about 15 mg/dl.

[0124] Blood (Haemoglobin)

[0125] In the presence of organic hydroperoxides and a chromogen, the peroxidase action of hemoglobin and myoglobin results in a green color. The intact erythrocytes are indicated by color spots in the reactive area, while the hemoglobin and myoglobin result in a uniform green color.

[0126] In the range of colors the following values are reported: 0 (negative), ca. 5-10, ca. 50, ca. 300 erythrocytes/L.

[0127] The concentrations are reported starting from about 5 erythrocytes/L.

[0128] Glucose

[0129] The test is based on the specific reaction glucose-oxidase/peroxidase.

[0130] In the range of colors the following values are reported: normal, 50, 100, 250, 500 and 1000 mg/dl. Glucose concentrations are reported starting from 40 mg/dl.

[0131] Ketone Bodies

[0132] The acetoacetic acid and acetone react with sodium nitroprusside in alkaline solutions, resulting in a compound of purple coloring (Legal test).

[0133] In the range of colors the following acetoacetic acid concentrations are reported: 0 (negative), 25 (+), 100 (++) and 300 (+++) mg/dl resp. 0 (negative), 2.5 (+), 10 (++) and 30 (+++) mol/l.

[0134] Urobilinogen

[0135] The test is based on the reaction of urobilinogen with a diazonium stable salt that results in a red azo coloring agent.

[0136] In the range of colors the following concentrations of urobilinogen are reported: normal, 2, 4, 8, 12 mg/dl and resp. normal, 35, 70.140, 200 mol/I.

[0137] Leukocytes

[0138] The granulocyte esterases dissociate an ester of carbonic acid heterocyclic. The fragment reacts with a diazonium salt and results in a purple coloring.

[0139] In the range of colors the following values are reported: 0 (negative), ca. 25, ca. 75, ca. 500 leukocytes/L. The concentrations are reported starting from 30 10-20 leukocytes/L.

[0140] Nitrites

[0141] The test is based on the principle of Griess reaction. Any pink coloring is to be interpreted as a positive result and indicates the presence of 105 organisms/ml of urine. The concentrations of nitrite are reported starting from 0.05-0.1 mg/dl.

[0142] PH

[0143] The test contains an indicator of mixing, able to differentiate clearly, in the values of pH 5 to 9, a range of colors ranging from orange, to yellow and cyan.

[0144] In the range of colors the following pH values are reported: 5, 6, 7, 8, 9.

[0145] Specific Weight/Density

[0146] The test is based on the color change of the reagent from blue-green to green-yellow, dependent on the concentration of ionic components in the urine. The test allows to determine the values of specific weight between 1.000 and 1.030.

[0147] In the range of colors the following concentrations are reported: 1.000, 1.005, 1.010, 1.020, 1.025 and 1.030.

[0148] Each of the above parameters corresponds to a different composition of the reaction pad.

[0149] The reactive components of the substrates, for the above parameters can therefore be the following: [0150] Ascorbic acid: 2,6-dichloro fenolindofenole (0.7%) [0151] Bilirubin: diazonium salt 3.1% [0152] Blood: tetramethylbenzidine dihydrochloride 2.0%, isopropilbenzolo, hydroperoxide 21.0% [0153] Glucose: glucose-oxidase 2.1%, peroxidase 0.9%, o-tolidine hydrochloride 5% [0154] Ketone bodies: sodium nitroprusside 2% [0155] Leukocytes: carbonic acid ester 0.4%, diazonium salt 0.2% [0156] Nitrites: tetraidrobenzochinolin-3-ol, 1.5%, sulfanilic acid 1.9% [0157] PH: methyl red 2.0%, bromothymol blue 10.0% [0158] Protein: bromophenol blue tetra 0.2% [0159] Specific Gravity: bromothymol blue 2.8% [0160] Urobilinogen: diazonium salt 3.6%

[0161] The present invention has hereto been described with reference to its preferred embodiments. It is to be mentioned that the technical solutions implemented in the embodiments described here as example, may advantageously be combined differently between them, to result to other embodiments afferent to the same inventive core and all however falling within the scope of protection of the claims hereafter reported.

[0162] In particular, the materials, methods and techniques used in the design and implementation of the present invention can be implemented by individual manufacturing companies based on their specific know-how, technology and operations, resulting in any case always included as part of what is expressed and claimed by the present invention.